Multiplexed transcriptome discovery of RNA-binding protein binding sites by antibody-barcode eCLIP

被引：0
|
作者：
Daniel A. Lorenz
Hsuan-Lin Her
Kylie A. Shen
Katie Rothamel
Kasey R. Hutt
Allan C. Nojadera
Stephanie C. Bruns
Sergei A. Manakov
Brian A. Yee
Karen B. Chapman
Gene W. Yeo
机构：
[1] Eclipse Bioinnovations,Department of Cellular and Molecular Medicine
[2] University of California San Diego,Institute for Genomic Medicine
[3] University of California San Diego,Stem Cell Program
[4] University of California San Diego,undefined
来源：
Nature Methods | 2023年 / 20卷
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摘要：
Ultraviolet crosslinking and immunoprecipitation (CLIP) methodologies enable the identification of RNA binding sites of RNA-binding proteins (RBPs). Despite improvements in the library preparation of RNA fragments, the enhanced CLIP (eCLIP) protocol requires 4 days of hands-on time and lacks the ability to process several RBPs in parallel. We present a new method termed antibody-barcode eCLIP that utilizes DNA-barcoded antibodies and proximity ligation of the DNA oligonucleotides to RBP-protected RNA fragments to interrogate several RBPs simultaneously. We observe performance comparable with that of eCLIP with the advantage of dramatically increased scaling while maintaining the same material requirement of a single eCLIP experiment.
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页码：65 / 69
页数：4
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